JPS62161798A - 3 alpha, 7 beta-dihydroxy-12-ketocholanic acid and its production - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides 3-alpha-7-peter dihydroxy-1
The present invention relates to a 2-ketocholanic acid compound and a method for producing the same.

Ursodeoxycholic acid (obtained by reducing 3-alpha-7-petadihydroxy-12-ketocholanic acid)
3-alpha-7-beta dihydroxycholanic acid) is known as a product of great therapeutic importance, as a gallstone solubilizer, for lowering the percentage of cholesterol in the blood, for lowering blood sugar. , as a diuretic,
It is also used as a promoter of lipid metabolism.

The currently used method for producing ursodeoxycholic acid is expressed by essentially the same reaction formula, that is, chenodeoxycholic acid is converted into 3α-hydroxy-7-ketocholanic acid (B), which is then hydrogenated. and ursodeoxycholic acid (C').

The known process consisting of the series of reactions described above has the following drawbacks. Compound CB is obtained by reduction of compound B.

A mixture is always obtained consisting of chenodeoxycholic acid and the compound chenodeoxycholic acid, from which it is difficult to remove chenodeoxycholic acid.

All methods proposed to date for separating mixtures of products A and B are industrially expensive and have limited efficiency.

The subject of the present invention is a novel compound and a process for its production which are useful for the production of highly pure ursodeoxycholic acid which is suitable for pharmaceutical use without any further purification steps and in particular is not contaminated with chenodeoxycholic acid. be.

The new compounds are prepared by a novel series of reactions starting entirely with cholic acid.

This new method essentially consists of the following steps.

1. 3α, 12α, dihydroxy 7 of cholic acid, i.e. 3α, 7α, 12α-trihydroxycholanic acid (I)
- Known methods to ketocholanic acid (If) (J, A;
C, S, 7 1 , 3935, 1 9 4 9 -
J, A, C, S, 72.5530.19
The steps necessary to prepare acid (II) from selectively oxidized cholic acid according to 50) are widely used commercially and are carried out by the commonly known methods described above.

2. 3α, 7β, of acid (II) by treating an aqueous solution of acid (■) with an alkali metal in 1-4C alcohol
Reduction to 12α-trihydroxycholanic acid (III) In practice in this reaction, acid (III) is formed in a limited amount of 10-15% relative to the 7α isomer, ie cholic acid (1).

3α, 7β, 12α trihydroxycholanic acid (4) is
by crystallization from a suitable solvent, or 3.7.1
It can be obtained in a pure state by esterifying part or all of 2.24 and purifying the ester obtained by crystallization from a suitable solvent. Acid (III)
is obtained from pure ester obtained by alkaline saponification.

However, for the present invention it is not necessary to obtain pure acid (I[). If position 3.7 is an acid and position 24 is 1
-40 alcohol, if properly esterified,
It can be used in the next step in its obtained state, that is, in its state containing cholic acid as an impurity. If necessary, these esterifications can be carried out continuously by conventional esterification methods.

product formula is a new compound.

[R is hydrogen or preferably an aliphatic acid group having 2 to 6 carbon atoms or an acid group selected from the group consisting of mono- or substituted benzoic acid, succinic acid, or glutamic acid; R' is hydrogen or an aliphatic group having 1 to 4 carbon atoms].

3.3α is esterified at positions 3 and 7.

7β,12α-trihydroxy-5β-cholanic acid was converted to 3α,
Oxidizes to 7β-dihydroxy-12keto-5β-cholanic acid.

弼 In the formula, R and R' are as defined above.

Excess hypochlorous acid is destroyed with bisulfite.

Free 3α,7β-dihydroxy 12keto-5β-cholanic acid is obtained by saponification with a strong alkaline dilute solution followed by acidification.

The free acid (IV) obtained in this way is mainly 3α,
Contains impurities consisting of 7α-dihydroxy-12keto-5β-cholanic acid and unreacted 3α-7β-12α-trihydroxycholanic acid. The product of formula (IV) is a new compound.

4.3α,7β-dihydroxy-12keto-5β-cholanic acid is treated with a silanizing agent such as bis-trimethylsilylurea, hexamethyldisilazane or bis-trimethylsilylacetamide in an organic solvent solution at a temperature of 30°C to 100°C. By processing, the entire tris-trimethylsilyl derivative is prepared and purified.

Surprisingly, compound (V) is only very slightly soluble in organic solvents. On the contrary, impurities and especially 3α, 7
α-dihydroxy-12keto-5β-furanic acid bathes very well in the same solvent.

The compound of formula (V) is also a new compound.

5. The trimethylsilyl group is removed by acid hydrolysis, preferably with hydrochloric acid, in an aqueous solution or an organic solvent according to the following formula:

(b) In this way, 3α,7β-dihydroxy-12-ketocholanic acid and its derivatives according to the present invention are obtained. Various esters of two human mixi groups and cal-xyl groups can be obtained by conventional methods.

6. The keto compound obtained in & is reduced to ursodeoxycholic acid as follows. 3α,7β-dihydroxy-12-keto- by heating to 200°C with hydrazine hydrate in the presence of an alkali group and triethylene glycol according to the Wurz-Kishchi method.
5β-cholanic acid is reduced to ursodeoxycholic acid.

Steps (5) and (6) may be reversed. Wurz-Killascher reduction can be carried out on tris-trimethylsilane derivatives, the trimethylsilyl group.

This is because it can be removed from an already reduced compound, ie, a tris-trimethylsilyl derivative of high purity ursodeoxycholic acid.

Essentially the same result can be obtained by interchanging the two steps.

The novel process according to the invention also has the advantage of using widely available starting materials.

It has the advantage that it can be converted to ursodeoxycholic acid in a few steps using a simple reaction with high selectivity. In addition, it has the advantage of directly producing pharmaceutically pure products without complicated and expensive purification processes.

Several examples are provided below so that the process of the present invention can be more easily reproduced. However, they do not limit the scope of the invention.

Example 1 Cholic acid (1) k selectively oxidized (J, A, C
, S. 71, 3955/1949: J, A, C, S.
72.5530/1950) obtained 3α, 1
2α, dihydroxy-7-ketocholanic acid, 100Iit
-Dissolve in secondary butyl alcohol 2000-. Heat the solution to boiling and add 100 II of sodium metal. Boiling is continued for 2 hours under reflux. Butyl alcohol is distilled and at the same time an equal amount of water is added. When all butyl alcohol has been removed, the mixture is cooled and acidified with 20% hydrochloric acid. The resulting precipitate is passed through the mouth, washed with water and dried. HPLC analysis shows 88.9% of 3α, 7β, 12α-trihydroxy-cholanic acid! A dry amorphous product 95II was obtained, consisting of 9.2% of -alic acid.

Acid titration 99.1% [α]D 71.2° (C-1% in dioxane) obtained in the previous step. ;-3α, 7β included as acidic impurities
, 200 m of benzene, 50 m of pyridine, and 50 d of acetic anhydride are added to 50 Ii of 12α-trihydroxycholanic acid. The mixture was allowed to react at 20°C to 25°C for 45 hours, then 50 m of water was added with stirring, and after a few minutes 200 m of water and 3
Add a mixture of 60-7% hydrochloric acid. The liquid phase is discarded and the organic phase is dehydrated to dryness.

The residue containing 3,7 diacetate was dissolved in 500% ethyl acetate, and 30 tons of 80% acetic acid and 15% sodium hypochlorite were added thereto.

The mixture 'f is stirred for 1 hour, the excess hypochlorite is destroyed with metabisulfite, and the ethyl acetate is washed with 500 ml of water.

Dehydrate the inorganic phase, dry it, and add 200 m of 10% NaOH.
Boil for 5 hours with g.

It is acidified with HCt and extracted with ethyl acetate.

A residue of 451 was obtained from the dehydrated and dried organic phase and was analyzed by HPLC.

50 g of the impure product obtained in the previous step are dissolved in 500 g of N,N-dimethylformamide.

Add 50 g of bis-trimethylsilyl urea.

The mixture is heated to 100° C., stirred at that temperature for 1 hour and then cooled.

3α,7β-bistrimethylsilyl ether-24 trimethylsilyl ester is crystallized, filtered through the mouth and washed with 50ml of N,N-dimethylformamide.

It is dried at 70°C in vacuum.

The properties of the obtained product of 60.9 are as follows: Molecular weight = 623.13 Melting point = 155° to 157°C % hydrozine hydrate 5o- and KOH 501 are added and the mixture is heated to about 200<0>C.

After the reaction is complete, cool it down, dilute it with 2000ml of water, and add HCl.
Make it acidic.

The product is filtered, washed with water and crystallized from ethyl acetate.

Cursodeoxycholic acid 2511 was obtained with the following properties.

Melting point = 202-205°C [α] = +60°±2 (C = 1% in dioxane)
Chenodeoxycholic acid impurity ≦1% Other impurities ≦0.3 Example 2 100 # of metallic potassium was boiled into 100 # of 3α, 12 α-nohydroxy-7-ketocholanic acid dissolved in 2000 # of n-butyl alcohol. and the mixture is heated under reflux for 2 hours.

Thereafter, all the n-butyl alcohol was distilled off while water was being supplied. After the distillation is finished, the mixture is diluted with 20% HCl
Acidify with water and separate the precipitated solids.

Solid (95,9)' (when subjected to i-HPLC analysis,
It can be seen that it consists of 3α, 7β, 12α-trihydroxycholanic acid with a 9° chain and cholic acid with a 10 chain. The properties of the mixture are similar to those in Example 1.

100 I of the mixture of 3α, 7β, 12α-trihydroxycholanic acid and cholic acid obtained in this step was dissolved in 150-pyridine, 100 g of succinic anhydride was added and the mixture was heated at 80° C. for 5 hours. do.

Cool to 0°C and add 100 g of water. Continue stirring for 2 hours. Add 1000 - of ethyl acetate and adjust - to 3-3.5 with HCt. Separate the aqueous phase.

The organic liquid obtained in this step contains 50-acetic acid and 50-
of 10% KBr water bath solution is added and the temperature is then adjusted to 20°C. While maintaining this temperature, a 150-15% sodium hypochlorite bath i' is added.

The mixture was stirred for 30 minutes and the product was distributed door to door.

Wash with 200W1t ethyl acetate and dry in a SO'C oven.

The dry product is boiled with 10001 nt of 10% NaOH for 2 hours, then cooled, 1000 ml of ethyl acetate is added and the mixture is acidified with 204 HO2.

The organic phase is separated and evaporated to 300 ml by volume.

Cool to 0°C and filter.

The product is washed with 100% ethyl acetate and dried in an oven at 80°C.

70y impure 3α,7β-dihydroxy-12-
Ketocholanic acid is obtained.

To 100 N of impure 3α,7β-dihydroxy-12-ketopranic acid obtained in this step, 100 td of acetonitrile, 700- of hexamethyldisilazane and 3
Add 00-trimethylchlorosilane. Heat under reflux for 1 hour, then cool. 3α,7β-bis-trimethylsilyl ether-24-) +7 methylsilyl ester crystallizes out, which is separated, washed with acetonitrile and dried under vacuum at 60°C.

The thus obtained tris-trimethylsilyl derivative 50
．． 9 is dissolved with stirring in 500 m of ethyl acetate and 100 m of 10% HCl for 10 minutes at a temperature of 50°C. The organic phase is separated and washed with 100 ml of water. Upon concentration, very pure 3α,7β-dihydroxy-12-keto-5β-furanic acid crystallizes out, which is filtered, washed with ethyl acetate and dried in the open at 80°C.

Characteristics: ・Molecular weight -406.5 ・Melting point 190℃ ・[α] = +108°±3 (C-1%, in dioxane) ・Total amount of impurities measured by chromatography ≦0.5% of 5011 Pure 3α,7β-dihydro-axy-12-ketocholanic acid was dissolved in 500-triethylene glycol, 50%
-80% of L dorazine hydrate and 501 (D KOH
and heat the solution at 200° C. for 1 hour.

After the reaction is complete, the mixture is cooled and diluted with 200-ml water.
Acidify with HCl. The product is separated by filtration and crystallized from ethyl acetate.

4511 extremely pure ursodeoxycholic acid is obtained, which has the following properties: Melting point 202-205°C [α), -+60'±2 (C-1% in dioxane) Impurities Total amount≦l Example 3 The same procedure as Example 1 was carried out except for the step involving the preparation of the tris-trimethylsilyl derivative. 3α, 7β- containing impurities dissolved in N-N-dimethylformamide 500-
Bis-trimethylsilyl-acetamide 50- is added to dihydroxy-12-ketocholanic acid 5011. The mixture is heated to 100° C. and held at that temperature for 1 hour with stirring, then cooled. The precipitate obtained after 1 hour at 100 DEG C. is recrystallized from N,N-dimethylformamide in the presence of bis-trimethylsilyl-acetamyl lo-.

It is passed through the mouth, washed with water and dried in vacuum at 70°C.

Tris-trimethylsilyl derivative of 3α,7β-dihydroxy-12-ketocholanic acid 61. f was obtained. Its characteristics are similar to those described in Example 1.

Claims (2)

[Claims] (1) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (In the formula, R is H, -Si(CH_3)_3 or an aliphatic or aromatic acid group, R' is H, -Si(CH_3)_3 or the number of carbon atoms 3α,7β-dihydroxy- of 1-4 alkyl)
12-ketocholanic acid compound. (2) Formula (II) ▲There are mathematical formulas, chemical formulas, tables, etc.▼ Starting from 3-α,12-α-dihydroxy-7-ketocholanic acid of (II), basically the following steps; (a) Compound By treating a solution of (II) in an aliphatic alcohol having 1 to 4 carbon atoms with an alkaline metal at the boiling temperature of the alcohol, formula (III) can be obtained. The compound 3-α,7-β,12-α-trihydroxy-
reduced to 5-β-cholanic acid; (b) the compound of formula (III) has 2 to 7 carbon atoms at the 3 and 7 positions;
6 fatty acids, benzoic acid, succinic acid or glutamic acid,
The corresponding group -OR (R is an acyl group) is obtained and esterified; (c) The esterification product of the previous step b is subjected to oxidation treatment with alkaline hypochlorite in acetic acid to form the formula ▲ mathematical formula, chemical formula , tables etc. ▼ Obtain the 12-keto derivative of (IV) (wherein R is an acyl group as defined in the previous step b); 3α,7β-dihydroxy-12 of formula (IV) consisting of −
A method for producing a ketocholanic acid compound.